Therapy-related acute myeloid eukemia/myelodysplastic syndrome (t-AML) is a complication that occurs in up to 10% of patients receiving chemotherapy with alkylating agents, topoisomerase II inhibitors, or radiation. This disease has increased in incidence as patients achieve longer remissions. T-AML remains refractory to current therapy and carries a uniformly fatal prognosis, with a median survival of 8-10 months. In this proposal, it is hypothesized that transformed hematopoietic cells have genetic mutations that drive leukemogenesis and can serve as therapeutic targets and biomarkers of disease. The specific aims are designed to test this hypothesis by sequencing the leukemic mRNA of t-AML patients using next-generation technology to identify genetic abnormalities, including potential gene fusions and mutations. Specific Aim 1 is the discovery and prioritization of molecular lesions in t-AML leukemia samples by sequencing all mRNAs in t- AML human samples. Specific Aim 2 is the validation of mutations by molecular techniques using the large number of t-AML samples available to us at the University of Chicago. Specific Aim 3 is to study the functional significance of mutations in vitro using a multitude of approaches dictated by the nature of the mutated gene products discovered. This information will allow the identification of pathways altered in t-AML, correlate these aberrations with clinical outcome, identify potential biomarkers, and generate candidate targets for leukemia therapeutics. PUBLIC HEALTH RELEVANCE: Therapy-related acute myeloid leukemia (t-AML) is a side-effect of chemotherapy that occurs in up to 10% of cancer survivors and does not have an effective treatment. Cutting-edge sequencing technology will be used to analyze the genome of t-AML cancer cells in this project. The goal of this proposal is to understand the genetics of t-AML and identify the cause and a treatment for this disease.